Ml Of Cerebrospinal Fluid Research Articles

P08.02 * IDENTIFICATION AND QUANTIFICATION OF CSF MALIGNANT CELLS BY THE CELLSEARCH(R) TECHNOLOGY IN PATIENTS WITH LUNG LEPTOMENINGEAL METASTASIS

INTRODUCTION: Diagnosis of lung leptomeningeal metastasis (LM) remains difficult. The usual diagnostic methods of cytomorphological assessment of cerebro-spinal fluid (CSF) and gadolinium-enhanced MRI lack both specificity and sensitivity. The Veridex CellSearch® technology is validated in detection of circulating tumor cells (CTC) in blood and in the follow-up of breast, prostate and colorectal cancer. Our aim was to adapt this technology for detection and quatification of tumor cells in the CSF of lung cancer patients with LM. METHODS: Patients with suspected lung LM from a french neuro-oncological center (Nancy) were prospectively included. The CSF samples were collected on traditional cytology tubes (1mL) and on CellSave® Preservative tubes (4 mL). RESULTS: 12 patients with clinical symptoms and/or radiological criteria suggestive of lung LM (5 at diagnosis/7 at recurrence) underwent CSF analysis with conventional cytology and with Veridex Cellsearch® technology. Conventional CSF cytomorphological analysis was positive in 5 patients (with a median of 13 tumor cells/mm3; range 10 to 25 tumor cells/mm3) whereas the assessment with Veridex Cellsearch® technology was positive in 12 patients. Quantitative analysis with the Veridex CellSearch® technology showed a median of 203 tumor cells/5mL of CSF (range 1 to 1500 tumor cells per 5 mL of CSF). CONCLUSIONS: In contrast to the current gold standard cytomorphological analysis, this new approach seems more sensitive and allowed a quantification of CSF tumor cells in lung LM. This methodology could be useful for earlier diagnosis of lung LM and for follow-up. A large prospective study is required.

Development of a method for analysis of ketamine and norketamine enantiomers in equine brain and cerebrospinal fluid by capillary electrophoresis.

Ketamine and norketamine are being transported across the blood brain barrier and are also entering from blood into cerebrospinal fluid (CSF). Enantioselective distributions of these compounds in brain and CSF have never been determined. The enantioselective CE based assay previously developed for equine plasma was adapted to the analysis of these compounds in equine brain via use of an acidic pre-extraction of interferences prior to liquid/liquid extraction at alkaline pH. CSF can be treated as plasma. With 100 mg of brain tissue and 0.5 mL of CSF or plasma, assay conditions for up to 30 nmol/g and 6 μM, respectively, of each enantiomer with LOQs of 0.5 nmol/g and 0.1 μM, respectively, were established and the assays were applied to equine samples. CSF and plasma samples analyzed stemmed from anesthetized patient horses and brain, CSF and plasma were obtained from anesthetized horses that were euthanized with an overdose of pentobarbital. Data obtained indicate that ketamine and norketamine enantiomers are penetrating into brain and CSF with those of ketamine being more favorably transported than norketamine, whereas metabolites of norketamine are hindered. More work is required to properly investigate possible stereoselectivities of the ketamine metabolism and transport of metabolites from blood into brain tissue and CSF.

Rapid diagnosis and discrimination of bacterial meningitis in children using gram probe real-time polymerase chain reaction.

In this study, we developed a method of simultaneous detection and discrimination of bacteria in cerebrospinal fluid (CSF) with gram probe real-time polymerase chain reaction (PCR). Our results showed 25 clinical strains representing 13 gram-positive and 12 gram-negative bacterial species. They were identified correctly with the corresponding gram probe. The standard curve showed that the amplification efficiency of templates with different concentrations of bacteria was almost the same with a potential detection limit of 10 colony-forming units/mL. A total of 482 children who were clinically suspected of bacterial meningitis were included in this study. A total of 1.0 mL of CSF was collected from every child and was subjected to gram probe-based PCR (GP-PCR), CSF culture, and CSF routine analysis. The positive rate of the GP-PCR array was (32/482, 6.64%) significantly higher than that of CSF culture (23/482, 4.77%). GP-PCR was proved to be an excellent technique for rapid and accurate diagnosis and discrimination of bacterial meningitis, and hence its use as a diagnostic tool in future seems very promising.

Cerebrospinal Fluid Correlates Of L-Asparaginase-Related Thrombotic Events

BackgroundThrombosis is a well-described complication of L-asparaginase (asp) treatment in patients with acute lymphoblastic leukemia (ALL), with an incidence of 5-10%. Although asp-associated thrombi can occur anywhere, most are venous and 50% of venous thromboses occur in the central nervous system (CNS); 18% of these are associated with cerebral infarction or stroke. Ninety percent of thrombotic events occur early in the treatment of ALL, during induction and consolidation, consistent with the predominant use of asp during these phases of therapy. Morbidity and mortality have been reported in as many as 50% of patients. Although the mechanism by which asp causes thromboses is thought to be inhibition of protein synthesis resulting in decreased levels of plasma coagulation factors, specific patterns of plasma coagulation factors (including decreases in antithrombotic proteins such as ATIII, protein S, C, and components of the fibrinolytic pathway) have not been good predictors of thrombosis. Risk factors for CNS thrombosis include older age at diagnosis and high risk (HR) disease. Although quantification of proteins in the cerebrospinal fluid (CSF) has been used to identify markers of other diseases, CSF proteomics has not been studied in ALL. We hypothesize that changes in some proteins in the CSF will occur after asp, will be more prominent in patients on HR ALL protocols than in those on standard risk (SR) protocols, and will anticipate CNS thrombosis. In this study, we pilot feasibility and describe serial proteomic analysis of CSF in newly diagnosed patients with ALL or lymphoblastic lymphoma (LL). MethodsSequential patients, ages 0-30 years, with B-cell ALL, T-cell ALL, or LL diagnosed August 2012-August 2013 at the University of North Carolina and treated according to risk stratified Children's Oncology Group protocols and the US Intergroup C10403 protocol for adolescents and young adults were offered participation. All patients were on a treatment protocol which utilized PEG-asp on Day 4 of therapy. Following consent approved by our Institutional Review Board, 1 mL of CSF was collected at the time of scheduled lumbar punctures (LP) on Days 0, 8, and 29 of induction. Samples were centrifuged at 1500rpm for 10 minutes within an hour of LP, stored in 50mcL aliquots at -800C. and transported on dry ice to the Duke Proteomics Core Facility. Because of cost ($1000/sample), 5 patients (15 samples) were selected for initial study, including the one patient who had a CNS thrombosis on day 22 of consolidation. Thawed batched duplicate samples were immunodepleted using a MARS-14 LC column (Agilent) and Agilent 1000 HPLC to remove high abundance plasma proteins, and quantitative MS analysis was performed in a label-free fashion using one-dimensional liquid-chromatography tandem mass spectrometry on a Synapt G2 HDMS system (Waters Corporation). Data Analysis was performed in Rosetta Elucidator v3.3 (Rosetta Biosoftware). ResultsSamples were obtained on each of 22 patients (11 male, 11 female). Characteristics of the 5 patient samples set chosen for analysis are shown in Table 1. 636 proteins were identifiable in each sample including several that have potential roles in coagulation. Thirteen proteins met a cut-off of ±1.5-fold change and p<0.05 for unpaired t-test for differential expression, including ADAMTS9, actin, protein Z-dependent protease inhibitor, SERPINA5 and SERPIND1.Table 1Patient CharacteristicsStudy #GenderAge (yrs)DiagnosisRisk2F11pre B ALLHR5F15mopre B ALLVHR7M8T cell LLstage III11M4T cell ALLIR13F8pre B ALLVHRIn the comparative analysis between the 4 patient sample sets and our index case with clot (subject 13), 28 proteins were differentially expressed in subject 13, on days 8 and 29, including lower levels of anti-thrombin III and plasminogen. ConclusionCSF proteomic analysis appears to be feasible and reproducible. These preliminary data suggest that sequential changes in CSF ATIII and plasminogen may predict CNS thrombosis. Reduced assay costs may be possible by targeting proteins of interest for analysis. Disclosures:No relevant conflicts of interest to declare.

Ultrasound-guided atlanto-occipital puncture for cerebrospinal fluid analysis on the standing horse

The atlanto-occipital site (AO) is convenient for retrieving an adequate volume and quality of cerebrospinal fluid (CSF) in the diagnosis of neurological disease in horses. However, general anaesthesia is not...

Detection and quantification of CSF malignant cells by the CellSearch® technology in patients with melanoma leptomeningeal metastasis

Melanoma is the most frequent solid tumor associated with leptomeningeal metastasis (LM). The usual diagnostic tools, that is, cytomorphological assessment of cerebro-spinal fluid (CSF) and gadolinium-enhanced MRI of the entire neuraxis both lack effectiveness. The CellSearch Veridex technology for the detection of circulating tumor cells (CTC) in blood was designed for the follow-up and prognosis of breast, prostate, colorectal, and lung cancer, which express EpCAM markers. We have previously adapted this technology to detect malignant cells in the CSF of breast cancer LM. Our objective here was to check if this technology would also allow the detection and the enumeration of CTC in the CSF of melanoma patients presenting with LM although melanoma does not express EpCAM markers. On the occasion of the intrathecal treatment of LM in 2 melanoma patients, 5 mL of CSF and 7.5 mL of blood were collected on CellSave Preservative Tubes and analyzed within 3 days after CSF sampling using a melanoma-dedicated kit. The CellSearch Veridex technology was then adapted to direct enrichment, enumeration, and visualization of melanoma cells in the CSF. CD146+, HMW-MAA+, CD34-, and CD45- cells with typical morphology could be observed and enumerated sequentially with reproducible results, corresponding to CSF melanoma cells (CSFMC). In contrast to the current gold standard cytomorphological analysis, this new approach allowed a precise quantification of CSFMC in all samples concomitantly analyzed. This methodology, established on a limited volume of sample and allowing delayed processing, could prove of great interest in the diagnosis and follow-up of melanoma patients with LM.

Brain death after Concorde positioning for supracerebellar-infratentorial approach: Unanswered questions and lessons learned

Brain death after Concorde positioning for supracerebellar-infratentorial approach: Unanswered questions and lessons learned

Hourly variability of cerebrospinal fluid biomarkers in Alzheimer's disease subjects and healthy older volunteers

Large hour-to-hour variability has previously been demonstrated in the cerebrospinal fluid (CSF) concentrations of Alzheimer's disease (AD) biomarkers amyloid β42 (Aβ42) and Aβ40 in healthy younger subjects. We investigated the within-subject variability over 36 hours in CSF Aβ and tau proteins, in older subjects and AD patients. Six patients with mild stage AD (59–85 years, Mini Mental State Examination (MMSE) 16–26) and 6 healthy older volunteers (64–77 years) received an intrathecal catheter from which, during 36 hours, each hour 6 mL of CSF was drawn. Concentrations of Aβ42, Aβ40, total tau, and phosphorylated tau were determined and the variability was analyzed. Within-subject variability within 3-hour periods was assessed as the coefficient of variation, which was comparable for these 4 biomarkers in controls (4.2%–4.6%) and AD (3.1%–5.8%). Variability over 12 hour periods was 5.3% to 9.5%. These findings suggest that CSF biomarker variability is relatively low in healthy older controls and AD patients. Furthermore, continuous sampling of CSF proved to be a useful and robust method, which may also be used to investigate AD pathogenesis and to evaluate pharmacotherapeutic interventions.

Hypertension intracrânienne et lupus

Introduction Idiopathic intracranial hypertension (IH) occurs most commonly in women and overweight subjects. It must be reported associated to general diseases, like systemic lupus erythematosus (SLE). Methods We report an observation of a patient with lupus complicated by glomerulonephritis and IH. Observation A 29 years old woman, without overweight, was followed for a SLE with skin and arthritic involvment. Four years after onset, a renal complication appeared with severe nephrotic syndrom. Six weeks after, bilateral papillar oedema was discovered, revealing an IH, as the patient was treated by oral steroids at 1 mg/kg/d and bimonthly intraveinous cyclophosphamide. The patient was completely asymptomatic. Brain MRI with veino-RMN was normal, without cerebral venous thrombosis. Lumbar punction showed an elevated opening pressure of 30,5 cmH 20 but with normal cerebrospinal fluid (CSF) contents. Evacuation of 30 mL of CSF and immunosuppressive treatment allowed symptoms regression. Discussion/Conclusion Twenty-seven cases of IH associated to SLE with nephritis have been reported in literature. Young women are more frequently involved with in half of cases a diffuse proliferative glomerulonephritis. Predisposing factors, like anaemia, must be associated. IH allows SLE diagnose in more than the third of the cases. Then, SLE has to be searched as an etiology of IH, in particular in non-obese patients and when nephritis is associated.

A minimally-invasive rat model of subarachnoid hemorrhage and delayed ischemic injury

Background:Double-injection models of subarachnoid hemorrhage (SAH) in rats are the most effective in producing vasospasm, delayed neurological deficits and infarctions. However, they require two large surgeries to expose the femoral artery and the atlanto-occipital membrane. We have developed a minimally-invasive modification that prevents confounding effects of surgical procedures, leakage of blood from the subarachnoid space and minimizes risk of infection.Methods:Rats are anesthetized and the ventral tail artery is exposed through a small (5 mm), midline incision, 0.2 mL of blood is taken from the artery and gentle pressure is applied for hemostasis. The rat is flipped prone, and with the head flexed to 90 degrees in a stereotactic frame, a 27G angiocath is advanced in a vertical trajectory, level with the external auditory canals. Upon puncturing the atlanto-occipital membrane, the needle is slowly advanced and observed for cerebrospinal fluid (CSF). A syringe withdraws 0.1 mL of CSF and the blood is injected into the subarachnoid space. The procedure is repeated 24 hours later by re-opening the tail incision. At 8 days, the rats are euthanized and their brains harvested, sectioned, and incubated with triphenyltetrazolium chloride (TTC).Results:Rats develop neurological deficits consistent with vasospasm and infarction as previously described in double-injection models. Cortical and deep infarctions were demonstrated by TTC staining and on histopathology.Conclusions:A minimally invasive, double-injection rat model of SAH and vasospasm is feasible and produces neurological deficits and infarction. This model can be used to study neuroprotective treatments for vasospasm and delayed neurological deficits following SAH, reducing the confounding effects of surgical interventions.

Diffusion Tensor Imaging Reveals Supplementary Lesions to Frontal White Matter in Idiopathic Normal Pressure Hydrocephalus

Idiopathic normal-pressure hydrocephalus (INPH) is associated with white matter lesions, but the extent and severity of the lesions do not cohere with symptoms or improvement after shunting, implying the presence of further, yet undisclosed, injuries to white matter in INPH. To apply diffusion tensor imaging (DTI) to explore white matter lesions in patients with INPH before and after drainage of cerebrospinal fluid (CSF). Eighteen patients and 10 controls were included. DTI was performed in a 1.5T MRI scanner before and after 3-day drainage of 400 mL of CSF. Regions of interest included corpus callosum, capsula interna, frontal and lateral periventricular white matter, and centrum semiovale. White matter integrity was quantified by assessing fractional anisotropies (FA) and apparent diffusion coefficients (ADC), comparing them between patients and controls and between patients before and after drainage. The significance level corresponded to .05 (Bonferroni corrected). Decreased FA in patients was found in 3 regions (P<.002, P<.001, and P<.001) in anterior frontal white matter, whereas elevated ADC was found in genu corpus callosum (P<.001) and areas of centrum semiovale associated with the precentral gyri (P<.002). Diffusion patterns in these areas did not change after drainage. DTI reveals subtle injuries-interpreted as axonal loss and gliosis-to anterior frontal white matter where high-order motor systems between frontal cortex and basal ganglia travel, further supporting the notion that motor symptoms in INPH are caused by a chronic ischemia to the neuronal systems involved in the planning processes of movements.

Fulminant ependymitis following intraventricular rupture of brain abscess

A 48-year-old man with a history of a penetrating brain injury was referred with a presumptive diagnosis of bacterial meningitis. Examination revealed a brain abscess in addition to meningitis. Blood and cerebrospinal fluid (CSF) cultures were negative for bacteria, and empirical IV antibiotic therapy with vancomycin (VCM) and meropenem was initiated. Despite initial improvement, however, his condition rapidly deteriorated into coma following intraventricular rupture of the abscess and hydrocephalus. Thereafter, an emergency ventriculostomy was performed and the abscess was evacuated. Bacterial cultures of the pus were negative. To manage the hydrocephalus, 150-200ml of CSF were drained daily. Intraventricular administration of VCM (20mg q.d.) was added to the IV antibiotic therapeutic regimen after surgery. Although the primary abscess rapidly decreased in size, ependymitis developed in the fourth ventricle. This new lesion, which resulted from CSF dissemination from the primary abscess, was refractory to treatment, and eventually disappeared after the intraventricular VCM dosage was increased from 20 to 30mg and continued for 30days. A possible reason for the development of fulminant ependymitis and why it was refractory to treatment despite the shrinkage of the primary lesion may be that physiological CSF flow from the lateral to the fourth ventricle was lost due to CSF drainage, and the stagnant CSF flow coupled with an insufficient VCM level in the fourth ventricle facilitated the rapid growth of pathogens. Although intraventricular antibiotic administration is efficacious for treating ruptured brain abscesses, it may be associated with the unexpected development of secondary lesions.

Proteomic investigations of the ventriculo-lumbar gradient in human CSF

Cerebrospinal fluid (CSF) is an ideal biological material in which to search for new biomarkers for improved diagnosis of neurological diseases. During a lumbar puncture between 5 and 15 mL of CSF are obtained. Previous studies have assessed the ventriculo-lumbar concentration gradient of a number of specific proteins. In the present study we took a proteomics approach to investigate the possible concentration gradient of a panel of proteins and peptides in the CSF of 16 patients with neurodegenerative diseases. Using two different mass spectrometry techniques, matrix assisted laser desorption ionization time of flight (MALDI-TOF) and surface enhanced laser desorption ionization time of flight (SELDI-TOF), we found that only one of the investigated proteins, apolipoprotein CI, was significantly decreased between the 1st and the 10th mL of CSF. Furthermore, we confirmed previous results showing a significant decrease in albumin concentration from the first to the last CSF aliquots. In conclusion, we found a significant gradient effect for only two of the measured proteins. However, a standardized procedure for CSF collection for diagnostic and research purposes is crucial to allow comparisons of results between patient groups and between laboratories. This is especially important since CSF is usually collected at several centres and variation in sampled CSF due to pre-analytical factors could complicate the interpretation of the results.

Diagnosis of cryptococcal and tuberculous meningitis in a resource-limited African setting

Cryptococcal meningitis (CM) and tuberculous meningitis (TBM) are common in HIV-infected adults in Africa and difficult to diagnose. Inaccurate diagnosis results in adverse outcomes. We describe patterns of meningitis in a Malawian hospital, focusing on features which differentiate CM and TBM with the aim to derive an algorithm using only clinical and basic laboratory data available in this resource-poor setting. Consecutive patients admitted with meningitis were prospectively recruited, clinical features were recorded and cerebrospinal fluid (CSF) was examined. A total of 573 patients were recruited, and 263 (46%) had CSF consistent with meningitis. One hundred and twelve (43%) had CM and 46 (18%) had TBM. CM was associated with high CSF opening pressure and low CSF leukocyte count. Fever, neck stiffness and reduced conscious level were associated with TBM. A diagnostic index was constructed demonstrating sensitivity 83%and specificity 79% for the differentiation of CM and TBM. An algorithm was derived with 92% sensitivity for the diagnosis of CM, but only 58% specificity. Although we demonstrate features associated with CM and TBM, a sufficiently sensitive and specific diagnostic algorithm could not be derived, suggesting that the diagnosis of CM and TBM in resource-limited settings still requires better access to laboratory tools.

Micrometer‐sized thread‐like and/or spherical particles in the first fraction of cerebrospinal fluid in patients with bipolar disorder

Scanning electron microscopy (SEM) is a powerful tool to identify pathogenic factors for which sensitive tests are lacking. The technique has been used to recognize structures in the cerebrospinal fluid (CSF) of patients with schizophrenia. The aim of this study was to use SEM to screen for potential particles in CSF in bipolar disorder. Fresh CSF samples from 56 euthymic bipolar patients, 31 bipolar I disorder and 25 bipolar II disorder, were compared to CSF samples from 20 controls. SEM of two portions of 200 microL filtered CSF was performed; the first 0.6 mL of CSF and the following 12 mL. The microscopic structures were identified and the quantity and patterns were rated by two independent researchers. Quantitative SEM examinations showed that of the 56 patients, 11 were free of any SEM structures in CSF, while 45 patients displayed morphological structures in the first 0.6 mL of CSF. By contrast, only 2 patients showed structures in the second CSF fraction drawn from the following mixed 12 mL of CSF. In total, 45 of the 56 patients had either thread-like, spherical, or both structures in the CSF, compared to none of the 20 controls. The identified particles in the first fraction of CSF have previously not been described in patients with bipolar disorder. Hypothetically, the amount of SEM structures in CSF, from none to many, might correlate to the degree of the alleged underlying disease processes in the central nervous system in patients with bipolar disease.

In-depth Exploration of Cerebrospinal Fluid by Combining Peptide Ligand Library Treatment and Label-free Protein Quantification

Cerebrospinal fluid (CSF) is the biological fluid in closest contact with the brain and thus contains proteins of neural cell origin. Hence, CSF is a biochemical window into the brain and is particularly attractive for the search for biomarkers of neurological diseases. However, as in the case of other biological fluids, one of the main analytical challenges in proteomic characterization of the CSF is the very wide concentration range of proteins, largely exceeding the dynamic range of current analytical approaches. Here, we used the combinatorial peptide ligand library technology (ProteoMiner) to reduce the dynamic range of protein concentration in CSF and unmask previously undetected proteins by nano-LC-MS/MS analysis on an LTQ-Orbitrap mass spectrometer. This method was first applied on a large pool of CSF from different sources with the aim to better characterize the protein content of this fluid, especially for the low abundance components. We were able to identify 1212 proteins in CSF, and among these, 745 were only detected after peptide library treatment. However, additional difficulties for clinical studies of CSF are the low protein concentration of this fluid and the low volumes typically obtained after lumbar puncture, precluding the conventional use of ProteoMiner with large volume columns for treatment of patient samples. The method has thus been optimized to be compatible with low volume samples. We could show that the treatment is still efficient with this miniaturized protocol and that the dynamic range of protein concentration is actually reduced even with small amounts of beads, leading to an increase of more than 100% of the number of identified proteins in one LC-MS/MS run. Moreover, using a dedicated bioinformatics analytical work flow, we found that the method is reproducible and applicable for label-free quantification of series of samples processed in parallel.

CT Ventriculography for diagnosis of occult ventricular cysticerci

Background:Neurocysticercosis is the most common parasitic infection of the central nervous system (CNS). Intraventricular lesions are seen in 7–20% of CNS cysticercosis. Intraventricular lesions can be missed by computed tomography (CT) and magnetic resonance imaging (MRI) as they are typically isodense/isointense to the cerebrospinal fluid. We present our experience with CT ventriculography to visualize occult cysts.Case Description:Two patients presented with hydrocephalus and suspected neurocysticercosis were evaluated with CT and MRI with and without contrast failing to reveal intraventricular lesions. CT-ventriculography was used: 10 ml of cerebrospinal fluid was drained from the ventriculostomy catheter, and 10 ml of iohexol 240 diluted 1:1 with preservative-free saline was injected through the ventriculostomy catheter. Immediate CT of the brain was performed. The first patient had multiple cysts located throughout the body of the left lateral ventricle. The second patient had a single lesion located in the body of the lateral ventricle. The CT-ventriculography findings helped in identifying the lesions and plan the surgical intervention that was performed with the aid of an endoscope to remove the cysts.Conclusions:Intraventricular neurocysticercosis is a common parasitic disease which can be difficult to diagnose. We used CT-ventriculography with injection of contrast through the ventriculostomy catheter in two patients where CT and MRI failed to demonstrate the lesions. This technique is a safe and useful tool in the imaging armamentarium when intraventricular cystic lesions are suspected.

Unusual Delayed Hydrocephalus After Bare Platinum Coil Embolization of an Unruptured Aneurysm -Case Report-

A 69-year-old woman developed hydrocephalus after the embolization of an incidentally detected unruptured large internal carotid artery aneurysm with bare platinum coils. Endovascular embolization resulted in near-total aneurysm occlusion. She complained of mild headache 18 hours after the procedure and magnetic resonance (MR) imaging performed on the 6(th) post-treatment day revealed wall enhancement and perianeurysmal brain edema. Follow-up MR imaging showed recanalization of the aneurysm and gradual ventricular enlargement. She presented with typical symptoms of hydrocephalus including disorientation, gait disturbance, and urine incontinence at 7 months post-embolization. We removed 30 ml of cerebrospinal fluid by lumbar tap, which improved her condition without symptom recurrence for 2 months. She did not require shunt placement. Post-interventional hydrocephalus is known in patients with unruptured aneurysms embolized with bioactive second-generation coils. This case shows that hydrocephalus can occur after aneurysm embolization with bare platinum coils without intracranial hemorrhage.

Determination of asialotransferrin in the cerebrospinal fluid with the HPLC method

Background: Identification of the content of asialotransferrin in the cerebrospinal fluid is a diagnostic method for childhood-onset ataxia and central nervous system hypomyelination (CACH), also known as vanishing white matter disease (VWM), and also for other types of CNS disorders. Methods: In our work, we have determined the value of the ratio of the asialo form of transferrin to the total transferrin in the CSF using the commercially used VariantTM Bio-Rad system for the determination of carbohydrate-deficient transferrin (CDT) in serum. The peak corresponding to the asialo form of transferrin was identified with electrophoresis with subsequent immunofixation and mass spectrometry (MALDI-TOF/TOF). Results: The intra-assay and inter-assay variations of the asialotransferrin value in CSF were 6.8% and 10.2%, respectively. Analysing CSF samples of 60 subjects (23 men aged 22–68 years and 37 women aged 18–77 years) with normal transferrin values and normal cytology as well as biochemistry parameters in the cerebrospinal fluid, and without apparent signs of neurological disorders, we have found the presence of 25.2 ± 8.2% asialotransferrin. Conclusion: Except for the need to obtain approximately 1.5 mL of cerebrospinal fluid and a tenfold concentrating of the sample, there is no need to conduct any modifications of the preparation procedure for the analytic sample and chromatographic separation normally used for serum samples. The HPLC method of asialotransferrin determination in CSF provides clinically useful results.

Improved Models of Mini Anion Exchange Centrifugation Technique (mAECT) and Modified Single Centrifugation (MSC) for Sleeping Sickness Diagnosis and Staging

Improved Models of Mini Anion Exchange Centrifugation Technique (mAECT) and Modified Single Centrifugation (MSC) for Sleeping Sickness Diagnosis and Staging